Abstract
An efficient protocol is described for the rapid in vitro plant regeneration of a medicinally important plant, Hygrophila spinosa through direct somatic embryogenesis from nodal explants excised from 4 week old aseptic seedlings. Somatic embryos differentiated directly from nodal explants on Murashige and Skoog (MS) medium supplemented with 1.0 µM 6- benzyladenine (BA) after 1 week of culture. The highest number (110.40 ± 3.55) of somatic embryos were recorded on MS medium supplemented with 1.0 µM BA and 0.5 µM NAA, growth regulator free half strength MS medium was found suitable for maturation and conversion of somatic embryos into complete plantlets. The well developed in vitro regenerated plantlets were successfully established in pots containing garden soil and grown in a greenhouse with 85% survival rate. The regenerants present an appearance similar to the seedlings obtained from normal seeds. The described method can be successfully employed for large scale multiplication and long term in vitroconservation of H. spinosa. Key words: Acanthaceae, Hygrophila spinosa, somatic embryogenesis, medicinal plant, conservation, micropropagation.
Highlights
We describe the first protocol for rapid in vitro plant regeneration of a medicinally important plant, H. spinosa through direct somatic embryogenesis from nodal explants followed by successful outdoor establishment of regenerated plants
All the cultures were incubated at 25 ± 2°C under 16 h photoperiod with a photosynthetic photon flux density (PPFD) of 50 μM m-2 s-1 irradiance provided by cool white fluorescent tubes with 60 - 65% relative humidity
Induction of embryogenic tissues was achieved when nodal explants excised from 4 week old aseptic seedlings were cultured on Murashige and Skoog (MS) medium supplemented with BA singly at different concentrations (Table 1)
Summary
High frequency induction of somatic embryos and plantlet regeneration from nodal explants of Hygrophila spinosa T. An efficient protocol is described for the rapid in vitro plant regeneration of a medicinally important plant, Hygrophila spinosa through direct somatic embryogenesis from nodal explants excised from 4 week old aseptic seedlings. Somatic embryos differentiated directly from nodal explants on Murashige and Skoog (MS) medium supplemented with 1.0 μM 6- benzyladenine (BA) after 1 week of culture. The well developed in vitro regenerated plantlets were successfully established in pots containing garden soil and grown in a greenhouse with 85% survival rate. The described method can be successfully employed for large scale multiplication and long term in vitro conservation of H. spinosa
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